Norbert Lukáč

Concentration of Copper, Iron, Zinc, Cadmium, Lead, and Nickel in Bull and Ram Semen and Relation to the Occurrence of Pathological Spermatozoa

Abstract In this study the concentration of copper, iron, zinc, cadmium, lead, and nickel in bull and ram semen and relation of these metals to spermatozoa morphology was investigated. Analysis by atomic absorption spectrophotometry showed that copper concentration was significantly higher (p < 0.0001) in ram semen in comparison with bull semen. The zinc concentration was higher in bull semen in comparison with ram semen. The iron and cadmium concentrations in the semen were similar. Higher concentration of lead was found in ram semen. Higher levels of nickel were found in ram semen in comparison with bulls. In bull semen 11.79 ± 4.88% of pathological spermatozoa was found. Higher occurrence of pathological spermatozoa was in ram semen (17.17 ± 3.76) in comparison with the semen of bulls. Separated tail, tail torso, and knob twisted tail were the most frequent forms of pathological spermatozoa in both species. Correlation analysis in bulls showed high positive relation between iron and zinc (r = 0.72), nickel and separated tail (r = 0.76), separated tail and tail torso (r = 0.71), tail torso and total number of pathological spermatozoa (r = 0.72), and between tail ball and total number of pathological spermatozoa (r = 0.78). In rams high positive correlation between cadmium and lead (r = 0.98), nickel and separated tail (r = 0.77), separated tail and total number of pathological spermatozoa (r = 0.69), knob twisted tail and retention of cytoplasmic drop (r = 0.78), and between knob twisted tail and other pathological spermatozoa (r = 0.71) was found. High negative correlation in ram semen was observed between copper and nickel (r = 0.71), copper and separated tail (r = 0.70), and between iron and tail torso (r = 0.67). The results suggest that the studied metals have a direct effect on spermatozoa quality.

Impact of oxidative stress on male fertility - a review.

Oxidative stress is a state related to increased cellular damage caused by oxygen and oxygen-derived free radicals known as reactive oxygen species (ROS). It is a serious condition, as ROS and their metabolites attack DNA, lipids and proteins, alter enzymatic systems and cell signalling pathways, producing irreparable alterations, cell death and necrosis. While small amounts of ROS have been shown to be required for several functions of spermatozoa, their excessive levels can negatively impact the quality of spermatozoa and impair their overall fertilising capacity. These questions have recently attracted the attention of the scientific community; however, research aimed at exploring the role of oxidative stress and antioxidants associated with male fertility is still at its initial stages. This review summarises the current facts available in this field and intends to stimulate interest in basic and clinical research, especially in the development of effective methods for the diagnosis and therapy of semen damage caused by oxidative stress.

Concentration of trace elements in human semen and relation to spermatozoa quality.

In this study the concentrations of trace elements such as lead, cadmium, iron, nickel, copper and zinc in the human semen (n = 47), occurrence of pathological spermatozoa, and correlations of these elements to pathological forms were investigated. For each sample of human spermatozoa at least 500 spermatozoa were evaluated. Metal contents were determined by the voltametric method and flame absorption spectrophotometry method. The concentrations of trace elements in human semen were: lead 1.49 ± 0.40 mg.kg− 1, cadmium 0.13 ± 0.15 mg.kg− 1, iron 2.59 ± 0.21 mg.kg− 1, nickel 0.40 ± 0.07 mg.kg− 1, copper 0.28 ± 0.06 mg.kg− 1, and zinc 153.93 ± 67.08 mg.kg− 1, respectively. The total percentage of pathological spermatozoa was 41.61 ± 9.80% with predominancy of broken flagellum, flagellum torso and separated flagellum. In relation to trace elements the analysis showed correlation between copper and lead (r = −0.47), nickel and iron (r = 0.36), lead and flagellum ball (r = −0.39), cadmium and large heads (r = 0.37) and between iron and other forms of pathological spermatozoa (r = −0.32). Results of this study describe possible effects of trace elements on the spermatozoa quality in normal human sperm.

Environmental levels of cadmium, lead and mercury in brown hares and their relation to blood metabolic parameters.

The purpose of this study was to examine concentrations of selected heavy metals in the liver and kidney of brown hares (Lepus europaeus). In addition, correlations between heavy metals and biochemical parameters in blood plasma were determined. The average concentrations of heavy metals (mmol/L) ± SD were as follows: liver: Pb 0.221 ± 0.189, Cd 0.160 ± 0.140, Hg 0.021 ± 0.030, kidney: Pb 0.115 ± 0.125, Cd 1.570 ± 1.103, Hg 0.030 ± 0.053. The average concentrations of biochemical parameters in the blood plasma were as follows: Ca 3.16 mmol/L, P 2.19 mmol/L, Mg 1.40 mmol/L, Na 148.71 mmol/L, K 8.12 mmol/L, glucose 6.56 mmol/L, total proteins 56.49 g/L, urea 5.00 mmol/L, total lipids 1.40 g/L, bilirubin 3.97 μ mol/L, cholesterol 1.53 mmol/L, aspartate aminotransferase (AST) 6.06 μ kat/L and alanine aminotransferase (ALT) 1.94 μ kat/L. Average levels of hormones (ng/mL) were as follows: testosterone 2.94, androstendiol 0.13, estradiol 501.59, progesterone 6.63, oxytocin 328.60. Tissue analysis showed an accumulation of lead, cadmium and mercury in the liver and kidney of brown hares. There were no significant correlations between levels of heavy metals in liver, kidney, and biochemical parameters.

Mercury-induced alterations in rat kidneys and testes in vivo

In this study effects of mercury administration on the kidney and testicular structure of adult rats were evaluated. Rats received mercury (HgCl2) in single intraperitoneal dose 20 mg HgCl2 (group A), 10 mg HgCl2 (group B) and 5 mg HgCl2 (group C) per kilogram of body weight and were killed after 48 hours following mercury administration. After the preparation of histological samples the results were compared with control group (K). In kidney decreased diameters of glomeruli and renal corpuscles, damaged tubules with affected quality of tubular cells and infiltration of interstitium were detected. Quantitative analysis demonstrated increased relative volume of tubules and renal corpuscles. Also the number of nuclei and glomeruli was increased in all experimental groups. The diameter of glomeruli and renal corpuscles was decreased. In testis undulation of basal membrane, dilatation of blood vessels in interstitium and occurrence of empty spaces in germinal epithelium were observed. Decreased relative volume of germinal epithelium, increased relative volume of interstitium and increased apoptosis occurrence suggest damaged interstitium and revealed occurrence of edemas. The relative volume of seminiferous tubules showed higher luminization. The number of nuclei was decreased in all experimental groups what is in positive relation with occurrence of empty spaces. Also other evaluated criteria demonstrated significant differences between control group and experimental groups. This study reports a negative effect of mercury on the structure and function of kidney and testes.

The impact of lead and cadmium on selected motility, prooxidant and antioxidant parameters of bovine seminal plasma and spermatozoa

The aim of this study was to investigate the effects of lead (Pb) and cadmium (Cd) content on basic motility characteristics (motility-MOT, progressive motility-PROG) as well as selected markers of the prooxidant-antioxidant balance (catalase-CAT, glutathione-GSH, malondialdehyde-MDA) in bovine seminal plasma and spermatozoa. Twenty five semen samples were collected from breeding bulls and used in the study. Motility analysis was carried out using the Computer Assisted Sperm Analysis (CASA) system. The samples were centrifuged, fractions of seminal plasma and spermatozoa were separated, lysates were prepared from the sperm cell fractions. Pb and Cd concentrations were determined by the voltametric method (ASV), antioxidants and MDA were analyzed by UV/Vis spectrophotometry. The analysis showed that the average concentration of Pb in the seminal plasma was 0.23 ± 0.02 μg/mL, while its amount in the sperm cells was significantly higher (0.41 ± 0.07 μg/mL; P < 0.05). The average Cd content in bovine seminal fractions was similar and non-significant: 0.09 ± 0.01 μg/mL in the seminal plasma and 0.11 ± 0.01 μg/mL in spermatozoa (P > 0.05). The correlation analysis revealed that both heavy metals were significantly negatively correlated with MOT and PROG (P < 0.05; P < 0.01; P < 0.001), CAT (P < 0.05; P < 0.01) as well as GSH (P < 0.05; P < 0.01) but significantly positively associated with MDA as the marker of lipid peroxidation (P < 0.05; P < 0.01). Moreover the samples were categorized in three quality groups (Excellent, Good, Moderate) according to their motility values. The lowest Pb, Cd and MDA concentrations but the best antioxidant characteristics were found in samples of the best quality, moderate quality samples exhibited the highest Pb, Cd and MDA content together with the worst antioxidant capacity. This study demonstrates that Pb and Cd are serious toxic elements, which are able to increase the risk of seminal oxidative stress development and a subsequent decrease of male fertility.

Reproductive toxicology of nickel – Review

The goal of this minireview is to summarize our current knowledge on the reproductive toxicity of soluble nickel salts. We made an attempt to present the most relevant data obtained from in vivo and in vitro experiments performed on mammals, mammalian primary cell cultures and cell lines. Nickel has been demonstrated to disturb the mammalian reproductive functions at several levels of regulation. The results of previous investigations indicate that the hormonal effects may play an important role in the reproductive toxicology of nickel both at the neuroendocrine and gonadal levels in the hypothalamic-pituitary-gonadal (HPG) axis. At the molecular level, it may be important that nickel may substitute certain other metals in metal dependent enzymes, leading to an altered protein function. It readily crosses the cell membrane via calcium channels and competes with calcium for specific receptors. Nickel can cross-link aminoacids to DNA, lead to formation of reactive oxygen species (ROS), moreover mimic hypoxia. These changes may lead to the activation of some signaling pathways, subsequent transcription factors and eventually to alterations in gene expression and cellular metabolism. These events are likely to be involved in the reproductive toxicity of nickel.

Concentration of Copper, Iron, Zinc, Cadmium, Lead, and Nickel in Boar Semen and Relation to the Spermatozoa Quality

Abstract The concentration of copper, iron, zinc, cadmium, lead, and nickel as well as its relation to spermatozoa quality was investigated. The semen samples were analyzed by atomic absorption spectrophotometry (AAS). The concentration of copper in boar semen was 1.64 ± 0.28 mg kg−1 and of iron 16.14 ± 10.35 mg kg−1. The concentration of zinc in boar semen reached an average value of 171.74 ± 64.72 mg kg−1 and the level of cadmium reached 0.01–0.16 mg kg−1 with the average value of 0.05 mg kg−1. The analysis of lead showed that the concentration of this element in boar semen was 0.02 ± 0.03 mg kg−1 and the average level of nickel was 0.06 ± 0.08 mg kg−1. The total percentage of pathological spermatozoa was 9.82 ± 1.47%. Detail analysis determined 3.18% of separated flagellum, 2.26% knob twisted flagellum, 0.88% flagellum torso, 0.85% flagellum ball, 0.42% broken flagellum, 0.23% retention of the cytoplasmic drop, 0.14% small heads, 0.03% large heads, and 1.83% forms other of pathological changes. Correlation analysis showed significant (p<0.05) positive correlation between copper and lead (r = 0.52). High correlation between small head and knob twisted tail (r = 0.67), small head and broken flagellum (r = 0.88) as well as between small head and total number of pathological spermatozoa (r = 0.73) was determined.

Bendiocarbamate induced structural alterations in rabbit thymus after experimental peroral administration

In this study histological structure of rabbit thymus after bendiocarbamate (2,3–isopropyledene–dioxyphenyl methylcarbamate) administration was studied. Bendiocarbamate was perorally administered for 90 days. At Day 3, 10, 20, 30, 60, 90 morphometric analysis was realized. Quantitative evaluation showed that in the control group thymus cortex forms 57.94 ± 7.10% and medulla 35.94± 7.38%. In almost all experimental groups significantly higher relative volume of cortex and lower relative volume of medulla was detected. Detail morphometric analysis found that the number of thymocytes per constant area and the diameter of tymocytes was decresed after bendiocarbamate administration. The number and diameter of reticular cells was not affected. Results of this study suggest negative effect of bendiocarbamate on the formation of thymus structures.

Seminal Concentration of Trace Elements in Fox and Relationships to Spermatozoa Quality

Concentrations of copper, zinc, iron, cadmium, lead, and nickel in the semen of foxes (Vulpes vulpes, n = 10), microscopic analysis of occurrence of pathological spermatozoa, and correlations of these elements with pathological forms were studied. Samples were analyzed by using an atomic absorption spectrophotometer. For analysis of pathological spermatozoa semen samples fixed with Hancocks solution and stained with Giemsa were prepared. For each fox at least 1000 spermatozoa were evaluated. The concentrations of copper, zinc, and iron in semen of foxes were found to be 2.16 ± 0.53 mg/kg, 13.09 ± 5.22 mg/kg, and 33.16 ± 24.36 mg/kg, respectively, on wet weight basis. Concentration of cadmium was low (0.07 ± 0.05 mg/kg). The levels of lead and nickel in the semen of foxes were 0.08 ± 0.06 mg/kg and 0.35 ± 0.24 mg/kg, respectively. The total percentage of pathological spermatozoa was 7.76 ± 1.33% with predominancy of knob twisted flagellum, separated flagellum, and broken flagellum. In relation to trace elements the analysis showed significant (p < 0.05) correlation between copper and lead (r = −0.85), copper and other forms of pathological spermatozoa (r = −0.72), zinc and broken flagellum (r = −0.69), iron and retention of cytoplasmic drop (r = 0.87), cadmium and separated flagellum (r = −0.68), and between cadmium retention of cytoplasmic drop (r = 0.87). This study detected significant effects of various trace elements in normal fox semen on the spermatozoa quality.